Objective:To investigate the mechanism of polygonatum and astragalus compound(PA)in inhibiting the progression of lung adenocarcinoma. Methods:CCK-8 assay was used to assess the inhibitory rate of proliferation in A549 and H1299 cells treated with PA at different concentrations and to calculate the half maximal inhibitory concentration(IC50).C57BL/6 mice(KRASG12D/+;TP53flox/flox)were treated with adenovirus carrying Cre enzyme via nasal inhalation to establish a mouse model of primary lung adenocarcinoma.The model mice were fed with PA-containing diet to directly observe the effect of PA on the lung adenocarcinoma tissue.Immunohistochemical staining was used to examine the pathological status of the lung tissue.Bioinformatics analysis indicated that PA affects the progression of lung adenocarcinoma through the apelin-peroxisome proliferator-activated receptor gamma coactivator 1-alpha(PGC1α)-mitochondrial brown fat uncoupling protein 1(UCP1).Real-time quantitative PCR and Western blotting analysis were used to study the effect of PA on the mRNA and protein expression levels of apelin-PGC1α-UCP1 signaling pathway related genes.An ATP detection kit and flow cytometry were used to evaluate the effect of PA on the ATP and mitochondrial ROS production,respectively,in A549 and H1299 cells.siUCP1 was used to silent the expression of UCP1 while Z160 was used to induce UCP1 overexpression in A549 and H1299 cells,and the changes in ATP and mitochondrial ROS production were examined to further investigate whether PA acts on apelin-PGC1α-UCP1 signaling pathway to affect the progression of lung adenocarcinoma. Results:PA could obviously inhibit the proliferation of A549 and H1299 cells with the IC50 values of 10.66 mg/mL for A549 cells and 9.66 mg/mL for H1299 cells.In the mouse primary lung adenocarcinoma model,PA could effectively inhibit the growth of tumor,downregulate apelin-PGC1α-UCP1 signaling pathway and inhibit the expression of lung adenocarcinoma-promoting gene UCP1.In A549 and H1299 cells,PA could significantly inhibit the expression of apelin,PGC1α and UCP1(P＜0.05),promote the production of ATP(P＜0.000 1)and ROS,restore mitochondrial oxidative phosphorylation,and inhibit aerobic glycolysis(P＜0.01).UCP1 silencing could increase the production of ATP(P＜0.01)and mitochondrial ROS and decrease the expression of key glycolysis enzymes hexokinase 2(HK2)and pyruvate kinase isozyme type M2(PKM2)(P＜0.05).Increasing the expression of UCP1 could reduce the ATP production(P＜0.01)and mitochondrial ROS generation in cells while increase the expression of HK2 and PKM2(P＜0.05).Treating cells with PA and Z160 simultaneously(PA+Z160)could reverse the inhibitory effect of PA on the ATP production and glycolysis of tumor cells(P＜0.05). Conclusion:PA can downregulate the apelin-PGC1α-UCP1 signaling pathway,inhibit mitochondrial uncoupling,restore mitochondrial oxidative phosphorylation,inhibit aerobic glycolysis,reverse the Warburg effect,and thus inhibit lung adenocarcinoma progression.

BACKGROUND: Sarcopenia is an age-related progressive skeletal muscle disorder involving the loss of muscle mass or strength and physiological function. Efficient and precise AI algorithms may play a significant role in the diagnosis of sarcopenia. In this study, we aimed to develop a machine learning model for sarcopenia diagnosis using clinical characteristics and laboratory indicators of aging cohorts. METHODS: We developed models of sarcopenia using the baseline data from the West China Health and Aging Trend (WCHAT) study. For external validation, we used the Xiamen Aging Trend (XMAT) cohort. We compared the support vector machine (SVM), random forest (RF), eXtreme Gradient Boosting (XGB), and Wide and Deep (W&D) models. The area under the receiver operating curve (AUC) and accuracy (ACC) were used to evaluate the diagnostic efficiency of the models. RESULTS: The WCHAT cohort, which included a total of 4057 participants for the training and testing datasets, and the XMAT cohort, which consisted of 553 participants for the external validation dataset, were enrolled in this study. Among the four models, W&D had the best performance (AUC = 0.916 ± 0.006, ACC = 0.882 ± 0.006), followed by SVM (AUC =0.907 ± 0.004, ACC = 0.877 ± 0.006), XGB (AUC = 0.877 ± 0.005, ACC = 0.868 ± 0.005), and RF (AUC = 0.843 ± 0.031, ACC = 0.836 ± 0.024) in the training dataset. Meanwhile, in the testing dataset, the diagnostic efficiency of the models from large to small was W&D (AUC = 0.881, ACC = 0.862), XGB (AUC = 0.858, ACC = 0.861), RF (AUC = 0.843, ACC = 0.836), and SVM (AUC = 0.829, ACC = 0.857). In the external validation dataset, the performance of W&D (AUC = 0.970, ACC = 0.911) was the best among the four models, followed by RF (AUC = 0.830, ACC = 0.769), SVM (AUC = 0.766, ACC = 0.738), and XGB (AUC = 0.722, ACC = 0.749). CONCLUSIONS: The W&D model not only had excellent diagnostic performance for sarcopenia but also showed good economic efficiency and timeliness. It could be widely used in primary health care institutions or developing areas with an aging population. TRIAL REGISTRATION Chictr.org, ChiCTR 1800018895.
Humans ; Aged ; Sarcopenia/diagnosis* ; Deep Learning ; Aging ; Algorithms ; Biomarkers

Objective To explore the safety and efficacy of endovascular treatment(EVT)in patients with acute posterior circulation ischemic stroke over 24 hours from onset.Methods This retrospective study retrospectively analyzed patients with acute posterior circulation ischemic stroke who received EVT in the Department of Neurology,First Hospital of Jilin University from June 2018 to June 2023.The patient's gender,age and other related demographic information were collected.The related examination results of patients were collected,including admission rapid blood glucose,admission systolic blood pressure,admission diastolic blood pressure.The related risk factors of stroke of patients were collected,including previous transient ischemic attack,hypertension,diabetes,atrial fibrillation,and history of drinking,smoking history,etc.;other related indicators were collected,including intravenous thrombolysis,tandem lesions,awakening stroke,baseline National Institutes of Health stroke scale(NIHSS)score,and baseline posterior circulation Alberta stroke program early CT score(pc-ASPECTS),collateral circulation grade of American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology(ASITN/SIR),onset to EVT puncture time,the trial of org 10172 in acute stroke treatment(TOAST)classification and outcome indicators(efficacy indicators included 90 days Modified Rankin scale[mRS]score≤3 after EVT,successful recanalization[extended cerebral infarction thrombolytic recanalization class 2b-3];security indexes included symptomatic intracranial hemorrhage[sICH]within 24 h after EVT and 90 d mortality after EVT).According to the symptom onset to EVT puncture time,the patients were divided into two groups:≤24h group and＞24h group.The patients with onset time＞24 h and those with onset time≤24 h were matched at a ratio of 1:1 by propensity score matching(PSM).All patients were divided into poor prognosis group(mRS score＞3)and good prognosis group(mRS score ≤ 3)according to mRS score at 90 days after EVT.Using univariate and multivariate Logistic regression was used to analyze the effect of onset time on clinical outcomes at 90 days after EVT.Results A total of 366 patients were enrolled in this study,including 284 males and 82 females.The median age was 61(55,68)years old,ranging from 25 to 91 years old.Before PSM,the＞24h group had lower prevalence of atrial fibrillation(2.02％[2/99]vs.9.74％[26/267],P=0.025)and lower baseline NIHSS score(10.0[5.0,19.0]vs.14.0[10.0,35.0],P＜0.01)and higher ASTIN/SIR collateral grade(P=0.018).After PSM,we did not find statistical difference between the two groups in baseline characteristics except for the onset to EVT puncture time.Before and after PSM,there was no significant difference in efficacy and safety between the onset time＞24 h group and the onset time ≤24h group.Univariate binary Logistic regression analysis showed that hypertension(OR,0.613,95％CI 0.391-0.942,P=0.025),intravenous thrombolysis(OR,3.235,95％CI 1.316-9.237,P=0.010),baseline NIHSS score(OR,0.975,95％CI 0.957-0.988,P＜0.01),baseline pc-ASPECTS(OR,1.281,95％CI 1.101-1.482,P=0.001)and sICH within 24 h after EVT(OR,0.070,95％CI 0.000-0.330,P＜0.01)were significantly correlated with prognosis.Gender,age,hypertension,baseline NIHSS score,intravenous thrombolysis,baseline pc-ASPECTS,ASTIN/SIR collateral grade,onset time＞24 h and sICH within 24 h after EVT were included in the multivariate binary Logistic regression analysis.The multivariate binary Logistic regression analysis showed that the onset time＞24 h was not associated with poor prognosis 90 d after EVT(aOR,1.635,95％CI 0.936-2.893,P=0.087).Conclusion EVT for acute posterior circulation ischemic stroke more than 24 hours after onset is feasible under strict imaging screening,and its safety and efficacy are similar to those in patients with onset under 24 hours.

Objective:To investigate the long-term clinical effect of deep brain stimulation (DBS) on patients with Parkinson's disease (PD) at Hoehn-Yahr stage 2.5-5 and the differences of clinical effect among patients at different Hoehn-Yahr stages.Methods:A total of 69 PD patients (7 at Hoehn-Yahr stage 2.5, 27 at stage 3, 31 at stage 4, and 4 at stage 5) accepted DBS in Department of Functional Neurosurgery, Affiliated Brain Hospital of Nanjing Medical University from May 2014 to December 2016 were selected for long-term follow-up observation at 60 months after DBS. Unified Parkinson's Disease Rating Scale (UPDRS) was evaluated in the "off" phase of the patients before DBS and the "off" phases of the drugs under DBS start-up and continuous treatment at 6, 12, 24, 36, 48 and 60 months after DBS, respectively; Parkinson's Quality of Life Questionnaire (PDQ-39) and levodopa equivalent dose (LED) data were collected before and 6, 12, 24, 36, 48 and 60 months after DBS. DBS parameters of the patients were collected at 12 months after DBS; the total stimulation power, single pulse average energy and large single pulse energy were calculated. The differences in UPDRS-III scores, PDQ-39 scores and LEDD in these patients before DBS and at different time points after DBS were compared; the differences in UPDRS-III and PDQ-39 scores at different time points after DBS and postoperative DBS parameters at 12 months after DBS in patients at different Hoehn-Yahr stages were compared.Results:Compared with those before DBS, the UPDRS-III scores, PDQ-39 scores and LED were significantly reduced at 6, 12, 24, 36, 48 and 60 months after DBS ( P<0.05). Significant differences in UPDRS-III scores were noted between patients at Hoehn-Yahr stage 5 and Hoehn-Yahr stage 2.5 and stage 3 at 12 and 24 months after DBS ( P<0.05); however, no significant differences in UPDRS-III scores were noted between patients at Hoehn-Yahr stage 5 and Hoehn-Yahr stage 4 at same time points ( P>0.05); and no significant differences in UPDRS-III scores were noted between patients at Hoehn-Yahr stage 5 and patients at Hoehn-Yahr stage 2.5, stage 3, and stage 4 at other time points ( P>0.05). No significant differences in PDQ-39 scores were noted between patients at different Hoehn-Yahr stages at 6, 12, 24, 36, 48, and 60 months after DBS ( P>0.05). No significant difference in total stimulation power, single pulse average energy or large single pulse energy was noted among patients at different Hoehn-Yahr stages at 12 months after DBS ( P>0.05). Conclusion:DBS is long-term effective in improving motor function and quality of life in PD patients at Hoehn-Yahr stage 2.5-5, and patients at Hoehn-Yahr stage 5 do not need higher DBS parameters to obtain similar clinical improved effects as patients at Hoehn-Yahr stage 2.5-4.

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has been widely used for genome engineering and transcriptional regulation in many different organisms. Current CRISPR-activation (CRISPRa) platforms often require multiple components because of inefficient transcriptional activation. Here, we fused different phase-separation proteins to dCas9-VPR (dCas9-VP64-P65-RTA) and observed robust increases in transcriptional activation efficiency. Notably, human NUP98 (nucleoporin 98) and FUS (fused in sarcoma) IDR domains were best at enhancing dCas9-VPR activity, with dCas9-VPR-FUS IDR (VPRF) outperforming the other CRISPRa systems tested in this study in both activation efficiency and system simplicity. dCas9-VPRF overcomes the target strand bias and widens gRNA designing windows without affecting the off-target effect of dCas9-VPR. These findings demonstrate the feasibility of using phase-separation proteins to assist in the regulation of gene expression and support the broad appeal of the dCas9-VPRF system in basic and clinical applications.
Humans ; Transcriptional Activation ; RNA, Guide, CRISPR-Cas Systems ; Gene Expression Regulation ; CRISPR-Cas Systems/genetics*

Objective:To explore the effectiveness and safety of endovascular treatment (EVT) for patients with acute anterior circulation ischemic stroke with symptom onset exceeding 24 h.Methods:In this retrospective cohort study, data were extracted from patients who underwent endovascular treatment for acute anterior circulation ischemic stroke at the First Hospital of Jilin University from February 2019 to April 2022. A total of 569 patients were included, with a mean age of 63 (54-70) years. Among them, 398 (69.9%) were male. The patients were divided into two groups based on symptom onset time:>24 h group and≤24 h group. Propensity score matching (PSM) was used to match the patients in a 1︰1 ratio between the>24 h group and the≤24 h group. Logistic regression was used to evaluate the impact of symptom onset time on outcome events.Results:Before PSM, compared with≤24 h group, the>24 h group had a younger age [56 (48, 64) vs. 64 (55, 70), Z=-3. 60, P<0.001]; lower proportion of prior atrial fibrillation [1.8% (1/57) vs. 21.1% (108/512), χ2=12.39, P<0.001]; lower proportion of wake-up stroke [7.0% (4/57) vs. 27.7% (142/512), χ2=11.54, P<0.001]; lower baseline NIHSS score [11.0 (7.5, 14.0) vs. 13.0 (10.0, 16.0), Z=-3.22, P<0.001]; and a higher American Society of Interventional and Therapeutic Neuroradiology/Society of Interventional Radiology(ASITN/SIR) grading ( P<0.001). After PSM, there were no significant differences in baseline characteristics between the two groups. There was no significant difference in the proportion of patients with a modified Rankin Scale (mRS) score≤2 at 90 days after surgery between the two groups (before matching: 42.0% vs. 40.4%, OR=0.745, 95% CI 0.407-1.362, P=0.339; after matching: 51.8% vs. 39.3%, OR=0.511, 95% CI 0.212-1.236, P=0.136). No significant differences were observed in the incidence of any safety outcomes between the>24 h group and the≤24 h group. Conclusion:For patients with acute anterior circulation ischemic stroke with symptom onset exceeding 24 h, EVT is feasible after strict radiological screening and has similar safety and effectiveness as for patients with symptom onset under 24 h.

Objective:To investigate the relationship between advanced glycation end products (AGEs) in the lens and type 2 diabetes mellitus.Methods:226 subjects were recruited between August 14 to September 14, 2018 from the Endocrinology Department of Central South University Xiangya Hospital, the Third Hospital of Changsha City, and the Fourth Hospital of Changsha City. The OGTT test, combined with clinical indicators, were used as the gold standard. Subjects were screened for type 2 diabetes using both the lens AGE fluorescence assay and the gold standard. Drawing the receiver operating characteristic (ROC) curve, we calculated the area under the curve (AUC) and its 95% CI and calculated the AGE for the diagnosis of type 2 diabetes. Sensitivity, specificity, Youden index, Kappa value, and its 95% CI, and the optimal cut-off value were determined according to the Youden index. Taking diabetes as the outcome indicator and AGE as the binary indicator, three logistic regression models were constructed. Stratified by age and sub-center, the differences between fasting blood glucose and 2 h postprandial blood glucose were compared between the AGE-negative and AGE-positive groups to determine the relationship between AGE and diabetes. Results:The area under the ROC curve was 0.86(95% CI: 0.81-0.91). According to the Youden index, the optimal cut-off point for AGE was 0.24. At this time, the sensitivity was 82.86(95% CI: 77.81-87.91), the specificity was 77.06(95% CI: 71.43-82.7), the Youden index was 59.92(95% CI: 53.36-66.49), the Kappa value was 79.62(95% CI: 74.22-85.02). Except for the 20-39-year-old group, the fasting blood glucose and 2 h postprandial blood glucose of the AGE-positive group in different age groups, different sub-centers, and the general population were higher than those of the AGE-negative group (all P<0.05). After adjusting for the confounding effects of age, gender, and sub-center (model 3), the relative risk of diabetes in the AGE-positive group was 11.75 times higher than the AGE-negative group (95% CI: 5.61-24.60), all with P<0.001. Conclusion:There was a high correlation between AGE in the lens and the risk of type 2 diabetes. When the cut-off point of AGE is 0.24, it had high sensitivity and specificity and could be used as a practical tool for early screening of type 2 diabetes.

The massive accumulation of β-amyloid （Aβ） in the brain is believed to be the first pathological mechanism of Alzheimer's disease （AD）， and the accumulation is mainly resulted from the overproduction and dysfunction in the clearance. Extensive and in-depth research has been carried out on AD. In addition to the drugs which are commonly used in clinical settings to improve cognitive function， Aβ monoclonal antibody aducanumab （Aduhelm） has been successfully marketed in the US， which may delay the progress of AD. Thus， it is a potential method for the treatment of AD to target Aβ， but it is expensive， with many adverse reactions and contraindications， which hinders the clinical promotion. Traditional Chinese medicine， featuring multiple components， multiple targets， multiple pathways， and high safety， can regulate the level of Aβ deposition in the brain， alleviate neurotoxicity， and prevent and treat AD by inhibiting the production and aggregation of Aβ and promoting the clearance in the brain. Berberine， gallic acid， osthole， scutellaria barbata flavonoids， Huannao Yicong decoction and Ditantang can promote α-secretase and inhibit the activity and expression of β- and γ-secretase， thus reducing production of Aβ. Baicalein， aloe-emodin， gallic acid， and curcumin can suppress the aggregation of Aβ， promote its depolymerization， and reduce the toxic effect of Aβ on nerve cells by interacting with the hydrophobic structure of Aβ and the H bond， salt bridge， and β-sheet that mediate the aggregation of Aβ. Curcumin and resveratrol can promote the expression of triggering receptor 2 in bone marrow cells of microglia and the migration and phagocytosis of Aβ in microglia. Bilobalide， Kaixinsan and curcumin can up-regulate the expression of encephalin-degrading enzyme and insulin degrading enzyme to promote Aβ degradation， and geniposide， dihydrotanshinone， dihydroartemisinin， and curcumin can degrade Aβ in cells by activating normal autophagy or inhibiting abnormal autophagy. Cycloastragenol， Danggui Shaoyaosan， Yizhi Fangdai formula and Linggui Zhugan decoction can promote the outflow of Aβ and inhibit the inflow of Aβ by improving the integrity and permeability of the blood-brain barrier （BBB）. Yizhi Fangdai formula and Xueshuantong can promote the polarization of aquaporin 4（AQP4）， allow Aβ to be cleared through the lymphatic system， and reduce the aggregation of Aβ in the brain， thereby relieving or preventing nerve cell damage and improving cognitive function. The above summary aims to provide more sufficient evidence and ideas for the clinical treatment of AD with traditional Chinese medicine.

Objective:To evaluate the value of 68Ga-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA)-fibroblast activating protein inhibitor (FAPI)-04 PET/CT on assessing different pathological grades in patients with renal fibrosis. Methods:A total of 25 patients (11 males, 14 females; age (39.3±13.9) years) diagnosed with renal fibrosis in the Affiliated Hospital of Southwest Medical University from September 2020 to August 2021 were retrospectively analyzed. All patients underwent renal puncture examination and 68Ga-DOTA-FAPI-04 PET/CT examination. The pathological results of kidney puncture were as the " gold standard" to divide the patients into mild fibrosis (Ⅰ), moderate fibrosis (Ⅱ), and severe fibrosis (Ⅲ). At the same time, 20 patients (10 males, 10 females; age (47.5±13.2) years) who underwent 68Ga-DOTA-FAPI-04 PET/CT examination showed no abnormal uptake of radioactivity in bilateral kidneys and no history of urinary system related diseases were enrolled as normal controls. Parameters including the maximum standardized uptake value (SUV max) of both kidneys, the mean standardized uptake value (SUV mean) of the liver, target/background ratio (TBR), glomerular filtration rate (GFR), and serum creatinine (Scr) were collected. Kruskal-Wallis rank sum test and Bonferroni correction method were used to compare the differences of SUV max, SUV mean, TBR and Scr among groups. One-way analysis of variance and least significant difference (LSD) t test were used to compare the difference of GFR among groups. The receiver operating characteristic (ROC) curve analysis was used to analyze the diagnostic efficacy of SUV max for the degree of renal fibrosis. Results:Of 25 patients, 22 had increased imaging agent uptake and the sensitivity of 68Ga-DOTA-FAPI-04 PET/CT in diagnosing renal fibrosis was 88%. The SUV max and TBR of patients with fibrosis grade Ⅰ, Ⅱ and Ⅲ were significantly higher than those of controls (SUV max: 4.40(3.30, 4.50), 5.90(4.28, 6.48), 8.50(7.50, 9.73) and 1.44(1.38, 1.68); TBR: 6.340±2.389, 8.097±1.420, 11.343±2.002 and 2.986±0.645; H values: 33.685, 32.368, all adjusted P<0.05 (Bonferroni correction method)). The Scr of patients with fibrosis grade Ⅰ and Ⅲ were significantly different (70.1(55.4, 92.5) and 174.1(161.4, 498.2) μmol/L; H=9.770, adjusted P<0.05 (Bonferroni correction method)). The liver SUV mean of patients with renal fibrosis grades Ⅱ and Ⅲ were significantly higher than that of controls (0.673±0.129, 0.751±0.170 and 0.514±0.142; H=15.609, both adjusted P<0.05 (Bonferroni correction method)). The GFR of patients with fibrosis grade Ⅲ had significant differences with grade Ⅰ and Ⅱ ((27.867±15.747), (87.756±31.657) and (63.160±29.556) ml/min; F=8.298, both P<0.05). ROC curve analysis showed that the area under curve was 0.946 7 (95% CI: 0.899 6-0.993 8, P<0.001). Conclusion:68Ga-DOTA-FAPI-04 PET/CT has a certain value in evaluating the degree of renal fibrosis, which can be used as a supplementary examination method for diagnosing renal fibrosis.

AIM: To investigate the effect and mechanism of hypoxia inducible factor-1α on intestinal mucosal barrier in sepsis. METHODS: SD rats were randomly divided into 4 groups: sham group, sepsis group, sepsis+HIF-1α stimulant (sepsis+DMOG group), sepsis+HIF-1α inhibitor (sepsis+Bay87-2243 group), 6 rats in each group. Sepsis model was established by cecal ligation and perforation (CLP). The levels of inflammatory markers IL-1β, IL-6, TNF-α, oxidative stress markers MDA and antioxidant factors SOD and CAT were detected by ELISA and the expression of HIF-1α in intestinal mucosa was detected by Western blot. The pathological damage of intestinal mucosa was detected by HE staining. RESULTS: Inflammatory factors, oxidative stress factors and HIF-1α were significantly up-regulated in septic rats (P<0.05). The contents of IL-1β, IL-6, TNF-α and MDA in plasma were significantly decreased by intraperitoneal injection of DMOG (P<0.05); the levels of SOD and CAT in plasma were increased (P<0.05), HIF-1α was up-regulated (P<0.05), and the pathological damage of intestinal mucosa was alleviated, with decreased Chiu's score (P<0.05). Oral administration of Bay87-2243 gave the opposite result. CONCLUSION: HIF-1α has a protective effect on intestinal mucosal injury in sepsis. The mechanism may be related to the alleviation of inflammatory response and inhibition of oxidative stress.